基于UHPLC-Q-TOF-MS/MS和网络药理学探究地龙抗疲劳作用机制  

Clarification of mechanism of Anti-fatigue Active Components Derived from Earthworms Based on UHPLC-Q-TOF-MS/MS and Network Pharmacology

作　　者：孟璇 王梦娇 赵天旸 张国忠 赵德龙 何希宏[1] 赵化冰 贾立明 贾京华 张黎明[1] MENG Xuan;WANG Mengjiao;ZHaO Tianyang;ZHANG Guozhong;ZHAO Delong;HE Xihong;ZHAO Huabing;JIA Liming;JIA Jinghua;ZHANG Liming(College of Biotechnology,Tianjin University of Science and Technology,Tianjin 300457,China;Tianjin Minsheng Bioengineering Co.Ltd.,Tianjin 301500,China)

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]天津民生生物工程股份有限公司,天津301500

出　　处：《现代食品科技》2025年第3期21-33,共13页Modern Food Science and Technology

基　　金：国家自然科学基金资助项目(22105145)。

摘　　要：基于超高效液相色谱四级杆-飞行时间串联质谱技术(Ultra High Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry/Mass Spectrometry, UHPLC-Q-TOF-MS/MS)分析与网络药理学探究地龙活性成分的抗疲劳作用机制。通过UHPLC-Q-TOF-MS/MS获得地龙化学成分信息,收集成分作用靶点、疲劳和抗疲劳的基因靶点取交集,进行基因本体(Gene Ontology, GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)富集分析。利用KEGG Mapper初步阐明抗疲劳机制,设计动物实验验证地龙抗疲劳功效。网络分析表明地龙抗疲劳主要集中在磷酸化的正向调节、核因子-κB(Nuclear Factor kappa-B, NF-κB)信号通路、5-羟色胺能突触、JAK-STAT通路和精氨酸和脯氨酸代谢通路等。动物实验结果表明,与空白组相比,地龙提取物中剂量组小鼠负重游泳实验时间由9.78 min延长至34.07 min(P<0.001),肝糖原含量显著升高,高剂量组为2 591.21 mg/g(P<0.001),高剂量组小鼠尿素氮和血乳酸含量分别降低至3.65 nmol/L和182.21 nmol/L(P<0.01)。综上,地龙通过多成分、多靶点、多途径的协同效应发挥抗疲劳作用,为后续的抗疲劳机制研究提供新的思路与方法。The anti-fatigue mechanism of the active constituents of earthworm was clarified using ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry/mass spectrometry(UHPLC-Q-TOF-MS/MS) and network pharmacology.UHPLC-Q-TOF-MS/MS was used to identify chemical constituents of earthworm extract.The targets of these constituents and fatigue and anti-fatigue associated genes were collected to identify the common targets.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed.KEGG Mapper was utilized to preliminarily elucidate the anti-fatigue mechanism,and animal experiments were performed to verify the anti-fatigue efficacy of earthworm extract.Network analysis showed that the anti-fatigue effects of earthworm extract mainly function via positive regulation of phosphorylation,NF-κB signaling pathway,serotonergic synapse signaling,JAK-STAT pathway,and arginine and proline metabolic pathways.As shown in the results of animal experiments,the time of weight-bearing swimming in mice in the middle-dose earthworm extract group was 34.07 min,significantly longer than that(9.78 min)of the CK group(P<0.001).Hepatic glycogen content in the high-dose earthworm extract group was 2591.21 mg/g,which was significantly higher than that in the CK group(P<0.001).Urea nitrogen and blood lactic acid contents in the high-dose earthworm extract group were 3.65 nmol/L and 182.21 nmol/L,respectively,which was significantly lower than those of the CK group(P<0.01).In conclusion,the anti-fatigue effect of earthworm extract was exerted through synergistic effects of multiple constituents,targets,and pathways,providing valuable insights and methods for further research into anti-fatigue mechanisms.

关 键 词：地龙 飞行时间串联质谱 网络药理学 抗疲劳 

分 类 号：R285[医药卫生—中药学]